Integrated airborne substance collection and detection system

ABSTRACT

A collection and detection system is configured as a detect to warn system in which the presence of specific types of particles are detected, and may or may not be identified. An air collection module intakes ambient air, detect the presence of one or more different types of airborne particles within the ambient air, and collect the airborne particles, such as within a fluid. A triggering mechanism is positioned to continuously monitor the airflow, to determine one or more characteristics of the airborne particles. If those measured characteristics match specific known characteristics, a trigger signal is generated. In response, a confirmation device performs a detection method on a fluid solution including the airflow particles to determine the presence of one or more different types of specific biological particles.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is related to commonly owned co-pending U.S.patent application Ser. No. (MFSI-00700), filed ______, entitled “AnIntegrated Airborne Substance Collection and Detection System”, and U.S.patent application Ser. No. (MFSI-00900), filed ______, entitled “AnIntegrated Airborne Substance Collection and Detection System”, whichare hereby incorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention relates to a method of and apparatus for collecting andanalyzing particulates. More particularly, the invention relates to thecollection and detection of airborne particulates.

BACKGROUND OF THE INVENTION

Bio-threat detectors are used to monitor the ambient air to detect thepresence of potentially harmful pathogens. In general, air is drawn intoa detection apparatus, or trigger, where the particulates in the air areevaluated. Airflow into the detection apparatus is typically generatedby a fan within the apparatus. The trigger continuously monitors the airand the individual molecules within a given airflow. Some triggers uselasers to scan the air path to interrogate the particles passingthrough. A harmless particle, such as a dust particle, can bediscriminated from a harmful particle, for example an anthrax spore,because each different type of particle reflects a different wavelengthof light. The laser light reflected off the passing particles is matchedto database of known harmful wavelengths. When a harmful wavelength isdetected, the trigger signals that a potential pathogen is present.However, the specific type of particle is not identified by the trigger.

A confirmation process is initiated once the trigger signals thepresence of a possible pathogen. During the confirmation process, theparticles that triggered the detection apparatus are identified.Conventionally, when the trigger goes off, the potential pathogen iscollected and taken to a lab where an analysis is performed. Multipletechniques are performed to identify the potential pathogen, eachtechnique is designed to identify a different type of pathogen,typically performed under the supervision of a lab operator. This is atime-consuming process requiring various pieces of test equipment, whichis impractical for real-time threat assessment. Such processes alsorequire the interaction of a human operator, which is costly and oftenefficient. Continuous monitoring and processing of potential pathogens,that is 24 hour a day coverage, requires multiple such human operatorsto cover the desired time frame.

SUMMARY OF THE INVENTION

Embodiments of a collection and detection system are directed to adetect to warn system in which the presence of specific types ofparticles are detected, and may or may not be identified. An exemplaryconfiguration of the integrated collection and detection system includesan air collection module, a confirmation device, and a control module.

In some embodiments, the air collection module is configured to intakeambient air, detect the presence of one or more different types ofairborne particles within the ambient air, and collect the airborneparticles, such as within a fluid. The air collection module includes atriggering mechanism and a fluid interface. The fluid interface isconfigured to receive ambient air, including airborne particles presenttherein, that is drawn into the collection and detection system and tocollect the airborne particles into a fluid solution. The triggeringmechanism is positioned to continuously monitor the airflow, and theairborne particles within the airflow, directed to the fluid interface.Through this interrogation of the ambient air, one or more opticalcharacteristics of the airborne particles are measured.

The optical characteristics are compared to known opticalcharacteristics to determine if one or more different types of specificbiological particles are present in the airflow. If it is determinedthat one or more different types of specific biological particles arepresent, than a trigger signal is generated.

In response to the trigger signal, the confirmation device confirms thepresence of the one or more different types of specific biologicalparticles. Confirmation is performed by attempting to detect the one ormore different types of biological particles within the fluid sample. Insome embodiments, an immuno assay is used to detect the one or moredifferent types of biological particles within the fluid sample.

In one aspect, a detection apparatus is configured to detect thepresence of one or more different types of particles. The detectionapparatus includes a first level detection device, a fluid interface,and a second level detection device. The first level detection device isconfigured to interrogate ambient air for the presence of one or moredifferent types of airborne biological particles, wherein the firstlevel detection device is further configured to generate a triggersignal in response to detecting one or more of the different types ofairborne biological particles. The fluid interface is configured toreceive airborne particles within the ambient air and to output a fluidsample including the particles. The second level detection device isconfigured to receive the fluid sample in response to the trigger signaland to confirm the presence of the one or more different types ofbiological particles using an immuno assay.

The detection apparatus can also include microfluidic circuitry tocouple the fluid interface to the second level detection device. Themicrofluidic circuitry can include a collection vessel to store thefluid sample. The microfluidic circuitry can also be configured to meterthe fluid sample into one or more portions and to distribute a firstportion of the fluid sample to the second level detection device. Thesecond level detection device can be configured to identify the one ormore different types of biologic particles using the immuno assay. Thebiologic particle can be a pathogen or a toxin. The second leveldetection device can include one or more capture devices each configuredto capture one or more different types of biological particles from thefluid sample. The second level detection device can also include anoptical detection module configured to optically detect the presence ofthe captured one or more different types of biological particles withinthe one or more capture devices. The detection apparatus can alsoinclude a control module configured to control the operation of thefirst level detection device, the fluid interface, and the second leveldetection device to enable the detection apparatus to functionautomatically. The detection apparatus can be configured to confirm thepresence of the one or more different types of biological particleswithin about 5 minutes from generating the trigger signal. The firstlevel detection device can include an air collection device configuredto intake the ambient air to be interrogated. The first level detectiondevice can also include a triggering mechanism configured to monitor theambient air taken in by the air collection device and to generate thetrigger signal. The triggering mechanism can include a light sourceconfigured to direct light onto the airborne particles and a lightcollector configured to measure one or more optical characteristics oflight impinging the airborne particles.

In another aspect, an autonomously functioning detection apparatus isconfigured to detect the presence of one or more different types ofparticles. The autonomously functioning detection apparatus includes afirst level detection device, a fluid interface, a second leveldetection device, and a control module. The first level detection deviceis configured to automatically interrogate ambient air for the presenceof one or more different types of airborne biological particles, whereinthe first level detection device is further configured to automaticallygenerate a trigger signal in response to detecting one or more of thedifferent types of airborne biological particles. The fluid interface isconfigured to automatically receive airborne particles within theambient air and to automatically output a fluid sample including theparticles. The second level detection device is configured toautomatically receive the fluid sample in response to the trigger signaland to automatically confirm the presence of the one or more differenttypes of biological particles. The control module is configured toprovide control signals to the first level detection device, the fluidinterface, and the second level detection device to enable the detectionapparatus to function autonomously.

In yet another aspect, a detection apparatus is configured to detect thepresence of one or more different types of particles. The detectionapparatus includes an air collection device, a triggering mechanism, afluid interface, a confirmation device, and a control module. The aircollection device is configured to intake ambient air including airborneparticles. The triggering mechanism is configured to interrogate theambient air for the presence of one or more different types of airbornebiological particles and to generate a trigger signal in response todetecting one or more of the different types of airborne biologicalparticles. The fluid interface is configured to receive airborneparticles within the ambient air and to output a fluid sample includingthe particles. The confirmation device is configured to receive thefluid sample in response to the trigger signal and to confirm thepresence of the one or more different types of biological particlesusing an immuno assay. The control module is configured to providecontrol signals to the air collection device, the triggering mechanism,the fluid interface, and the confirmation device to enable the detectionapparatus to function autonomously.

In still yet another aspect, a method of detecting the presence of oneor more different types of biological particles is described. The methodincludes collecting ambient air including airborne particles, measuringone or more optical characteristics of the airborne particles, comparingthe measured one or more optical characteristics to known opticalcharacteristics associated with one or more types of biologicalparticles, generating a trigger signal if the one or more of themeasured one or more optical characteristics match one or more of theknown optical characteristics, converting the airborne particles to afluid sample, and analyzing the fluid sample according to an immunoassay to confirm the presence of the one or more specific types ofbiological particles.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates an exemplary network configuration including multiplecollection and detection systems.

FIG. 2 illustrates an exemplary functional block diagram of a firstembodiment of the integrated collection and detection system.

FIG. 3 illustrates an exemplary block diagram of the control module.

FIG. 4 illustrates an exemplary schematic diagram of the archive module.

FIG. 5 illustrates an exemplary schematic diagram of the toxin captureand detection module.

FIG. 6 illustrates an exemplary schematic diagram of the lysis andcapture module.

FIG. 7 illustrates an exemplary schematic diagram of the metering andthermal cycling module.

FIG. 8 illustrates an exemplary schematic diagram of the opticaldetection module.

FIG. 9 illustrates an exemplary automated process performed by the firstembodiment of the particle collection and detection system.

FIG. 10 illustrates an exemplary functional block diagram of the secondembodiment of the integrated collection and detection system.

FIG. 11 illustrates an exemplary automated process performed by thesecond embodiment of the particle collection and detection system.

FIG. 12 illustrates an exemplary functional block diagram of the thirdembodiment of the integrated collection and detection system.

FIG. 13 illustrates an exemplary automated process performed by thethird embodiment of the particle collection and detection system.

Embodiments of the integrated particle collection and detection systemare described relative to the several views of the drawings. Whereappropriate and only where identical elements are disclosed and shown inmore than one drawing, the same reference numeral will be used torepresent such identical elements.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

Embodiments of the present invention are directed to a fully integratedand autonomous, collection and detection system configured to monitorthe ambient air for specific particles, such as pathogens. In someembodiments, the collection and detection system is configured as anintegrated cartridge. In some embodiments, the collection and detectionsystem is configured as a fully autonomous system. An air collectorcaptures airborne particles and outputs a fluid sample including thecaptured particles in a fluid solution. The collection and detectionsystem includes a control module configured to control the processing ofthe fluid sample such that detection of one or more types of particlesis fully automated within the integrated cartridge. The types ofparticles to be processed and detected include, but are not limited to,cells, bacteria, viruses, nucleic acids, toxins, and other pathogens. Ifone or more specific types of particles are detected, a system alarm istriggered. In some embodiments, the system alarm is an alarm signalwhich is transmitted over a communications network to either a local orcentral monitoring location. More than one collection and detectionsystem can be coupled to the network and monitored by the centralmonitoring location. In other embodiments, the system alarm is an audioand/or visual signal generated by the collection and detection systemitself.

FIG. 1 illustrates an exemplary network configuration including multiplecollection and detection systems 10. Each collection and detectionsystem 10 can be operated independently, or networked to a remotemonitoring location, as is illustrated in FIG. 1. The monitoringlocation can be local, as in the local monitoring point 40, orcentralized, such as the central monitoring point 50. As shown in FIG.1, each collection and detection system can operate independently, canbe coupled to a local monitoring point, which in turn can be coupled toa central monitoring point, or can be coupled to the central monitoringpoint. The collection and detection system 10 is coupled to the localmonitoring point 40 or the central monitoring point 50 via anyconventional network 60. Network connectivity also enables remotecontrol signal to be provided to the collection and detection system 10.

A first embodiment of the integrated collection and detection system isdirected to a detect to treat system in which specific particles areidentified. FIG. 2 illustrates an exemplary functional block diagram ofthe first embodiment of the integrated collection and detection system.The integrated collection and detection system 10 includes a controlmodule 12, an air collection module 14, a distribution module 16, anarchive module 18, a lysis and capture module 20, a toxin capture anddetection module 22, a solutions module 24, a solutions module 26, awaste module 28, a metering and thermal cycling module 30, a solutionsmodule 32, and an optical detection module 34. Fluid is directed betweenmodules and within each module using microfluidic pathways and valves,also referred to as microfluidic circuitry.

The air collection module 14 is configured to intake ambient air andcollect airborne particles within the air. Air is collected for apredetermined time frame, after which the collected particles are elutedinto a liquid sample which is output from the air collection module 14.The fluid sample output from the air collection module 14 includes afluid and particle solution.

The distribution module 16 meters and distributes the fluid sampleoutput from the air collection module 14. The fluid sample is meteredand distributed according to predetermined ratios. A first portion ofthe fluid sample is directed to the archive module 18, a second portionto the lysis and capture module 20, and a third portion to the toxincapture and detection module 22. In one embodiment, a syringe pump isused as part of the microfluidic circuitry to meter the fluid sample. Asyringe pump is adaptable for changing applications, such as changingthe distribution ratio from one application to the next. In anotherembodiment, a reservoir with drain holes is included as part of themicrofluidic circuitry. The location of each drain hole corresponds to adesired distribution ratio. A valve is coupled to the drain line of eachdrain hole to control the collection and distribution of the fluidsample between runs. Such a configuration is appropriate where thedistribution ratio is fixed, as the location of the drain holes is afixed specification. In yet another embodiment, aspects of a fixed ratioconfiguration, such as the reservoir with drain holes, is combined withaspects of the adjustable ratio configuration, such as the syringe pump.It is understood that other microfluidic circuit configurations can beused to meter and distribute the fluid samples for both fixed andvariable distribution ratios.

The archive module 18 is configured to store one or more fluid samples.The fluid samples are stored for later analysis and/or confirmation, ifnecessary. The lysis and capture module 20 is configured to perform alysis, purification, and concentration process on the fluid samplereceived from the distribution module 16. Lysis is performed on cellswithin the received fluid sample that are capable of being lysed. Lysisis performed using sonication. Alternatively, any conventional lysismethod can be used. Once the cells are lysed, the resulting nucleicacids are purified and concentrated to be sent to the metering andthermal cycling module 30. The solutions module 24 provides solutionsused during the lysis, purification, and concentration steps performedin the lysis and capture module 20. For example, the solutions module 24includes wash solutions and elution buffers.

The metering and thermal cycling module 30 receives the concentratedfluid sample from the lysis and capture module 20. The received fluidsample is metered and distributed into a predetermined number ofcollection vessels. The metering and thermal cycling module 30 iscoupled to the solutions module 32 to receive mixing solution that ismetered and distributed to each collection vessel such that acombination of concentrated fluid sample and mixing solution aretemporarily stored in each collection vessel. Each collection vessel iscoupled to a corresponding thermal cycling chamber to successively heatand cool the combined solution. In this manner, the fluid sample andmixing solution combination within each collection vessel undergoes athermal cycling process within the thermal cycling chambers to amplifyany nucleic acids present in the fluid sample. Any number of thermalcycles can be performed. This amplification process can be repeated, forexample a pre-amplification step and an amplification step can beperformed.

The amplified fluid sample from each thermal cycling chamber issuccessively output from the metering and thermal cycling module 30.Each amplified fluid sample output from the metering and thermal cyclingmodule 30 is interrogated by the optical detection module 34. Ingeneral, any conventional luminescence detection technology can beapplied to perform biological detection. The raw data obtained by theoptical detection module 34 is provided to the control module 12, whereit is used to determine the presence of one or more types of nucleicacids. If a nucleic acid is detected, the control module 12 generates analarm signal. Alternatively, the raw data collected by the opticaldetection module 34 is sent to a remote location, such as the centralmonitoring point 50 (FIG. 1) for analysis.

The toxin capture and detection module 22 is configured to capturetoxins present in the fluid sample received from the distribution module16. The toxin capture and detection module 22 is also configured todetect the presence of any captured toxins using any conventionalluminescence detection technology. The raw data obtained by the toxincapture and detection module 22 is provided to the control module 12,where it is used to determine the presence and identity of one or morespecific types of toxins. If a specific toxin is detected, the controlmodule 12 generates an alarm signal. Alternatively, the raw datacollected by the toxin capture and detection module 22 is sent to aremote location, such as the central monitoring point 50 (FIG. 1) foranalysis. In one embodiment, the toxin capture and detection module 22includes an optical detection device configured to measure one or morecharacteristics of any captured toxin. The solutions module 26 providessolutions used during the toxin capture steps performed in the toxincapture and detection module 22. For example, the solutions module 26includes wash solutions and antibody solutions.

The collection and detection system 10 is configured to be re-used suchthat successive fluid samples output by the air collection module 14 areprocessed. As such, the distribution module 16, the lysis and capturemodule 20, the toxin capture and detection module 22, the metering andthermal cycling module 30, and all interconnecting microfluidiccircuitry including the microfluidic circuitry coupling the metering andthermal cycling module 30 and the optical detection module 34 aredecontaminated between cycles. Various solutions are used to perform therinse and wash steps during decontamination, these solutions areincluded in the solutions module 24 and the solutions module 26.

The control module 12 is coupled to each module to control operation ofthe collection and detection system 10. Such control enables completeautomation of the collection and detection process, without need ofhuman intervention. The control module 12 is also configured to analyzethe raw data provided by the toxin capture and detection module 22 andthe optical detection module 24, and to generate any appropriate alarmsignals. In response to an alarm signal, the control module 12 initiatesa localized audio and/or visual alarm and/or transmits a notificationsignal to a networked local monitoring location or a centralizedmonitoring location.

The analyzed fluid samples, elution buffers, mixing solutions, rinses,washes, purged archive samples, and other solutions related to theprocessing of fluid samples and subsequent decontamination of thecollection and detection system 10 are directed to the waste module 28.Alternatively, fluid samples analyzed and subsequently output by thetoxin capture and detection module 22 and the optical detection module34 can be archived, either in the archive module 18, or a supplementalarchive module (not shown). The embodiments of the particle collectionand detection module 10 described above include three solutions modules.Alternatively, one or more of the solutions modules 24, 26, and 32 canbe combined, or more than three solutions modules can be used.

The system implementation illustrated in FIG. 2 is for illustrativepurposes. The microfluidic circuitry and module nature of the integratedcollection and detection system provides flexibility and extensibilityto interconnect and configure the modules, and associated sub-modularcomponents, into any desired combination. For example, the fluid samplecan be metered into additional portions, and each portion can be furthersub-divided into smaller portions. These portions can be distributed toany one of a multitude of fluid processing pathways, including the fluidpathway through the lysis and capture module 20 and the metering andthermal cycling module 30, the fluid pathway through the toxin captureand detection module 22, and any other fluidic pathway configuredaccording to one or more of the modules and/or sub-modules describedabove. As an additional example, a lysis module similar to the lysiscomponent in the lysis and capture module 20 can be added prior to thetoxin capture and detect module 22 to lyse cells prior to delivering thefluid sample to the toxin capture and detect module 22. Similar parallelpathways can also be configured such that a portion of the fluid sampleis received unlysed by the toxin capture and detect module 22, andanother portion of the fluid sample is first lysed by a lysis componentand then the lysed sample is delivered to another toxin capture anddetect module. Additionally, the specific configurations described foreach of the modules is for exemplary purposes. The microfluidiccircuitry and constituent components of each module can be adapted intoany number of configurations to perform the described functionality.

FIG. 3 illustrates an exemplary block diagram of the control module 12.The control module 12 includes a processor 122, a host memory 124, amass storage 126, and an I/O interface 130, all coupled via a system bus128. The mass storage 126 can include both fixed and removable mediausing any one or more of magnetic, optical or magneto-optical storagetechnology or any other available mass storage technology. The hostmemory 124 is a random access memory (RAM). The processing module 122 isconfigured to control the operation of the collection and detectionsystem 10. The I/O interface 130 includes a user interface and a networkinterface. In some embodiments, the user interface includes a display toshow user instructions and feedback related to input user commands. Thenetwork interface includes a physical interface circuit for sending andreceiving data and control communications over a conventional network,such as to a local or centralized monitoring location.

FIG. 4 illustrates an exemplary schematic diagram of the distributionmodule 16 coupled to the archive module 18. In this exemplaryconfiguration, the distribution module 16 includes a metering module162, a wash syringe 164, a syringe pump 166, and a peristaltic pump 168coupled together via microfluidic circuitry including valves 169-180.The archive module 18 includes five archive chambers 181-185 coupled tothe distribution module 16 via microfluidic circuitry including thevalves 186-195.

The fluid sample provided by the air collection module 14 is stored inthe metering module 162. In general, the amount of fluid sample providedby the air collection module 14 is an inconsistent amount. In oneembodiment, the collection and detection system 10 is configured toprocess a specific amount of fluid sample, in this case 10 ml. As such,a first step is to remove excess fluid sample from the metering module162. As applied to the configuration of FIG. 4, any excess fluid sampleis removed from the metering module 162 by opening the valve 173 and thevalve 179, which enables any excess fluid sample to flow to waste.Remaining is the specific amount of fluid sample in the metering module162.

Each archive chamber 181-185 is configured to store a predeterminedamount of fluid sample. In one embodiment, each archive module 181-185is configured to store 1 ml. This predetermined amount of fluid sampleis metered from the metering module 162 and delivered to one of thearchive chambers 181-185 by opening the valves 174 and 169 and thevalves corresponding to the archive chamber, such as the valves 186 and191 for archive chamber 181, turning on the peristaltic pump 168 in afirst direction, which forces air from the vent at the valve 169 intothe metering module 162. This pressurizes the metering module 162thereby forcing the fluid sample within through the open valves 174 and191 and into the archive module 181.

One archive chamber stores the fluid sample for the current cycle, andthe remaining four archive chambers store the fluid samples from theprevious four cycles. During the next cycle, the oldest fluid sample inthe archive is removed and replaced by the next fluid sample. Forexample, during a first cycle, a first fluid sample is received from thedistribution module 16 and stored in the archive chamber 181. During asecond cycle, a second fluid sample is received and stored in thearchive chamber 182. During a third cycle, a third fluid sample isreceived and stored in the archive chamber 183. During a fourth cycle, afourth fluid sample is received and stored in the archive chamber 184.During a fifth cycle, a fifth fluid sample is received and stored in thearchive chamber 185. During a sixth cycle, the first fluid sample storedin the archive chamber 181 is first purged to waste. To purge the fluidsample from the archive chamber 181, the valves 172, 186, 191, and 179are opened and the peristaltic pump 168 is run in a second direction,which forces air from the vent at the valve 172 into the archive chamber181. This pressurizes the archive chamber 181 thereby forcing the fluidsample within through the open valves 191 and 179 to waste. The valves172, 186, 191, and 179 are then closed and the archive chamber 181 isthen washed using solution provided via the wash syringe 164. The sixthfluid sample is then provided from the distribution module 16 to theempty archive chamber 181. Subsequent fluid samples are stored in asimilar manner such that the most recent five fluid samples are archivedin the archive module 18.

After the first portion of the fluid sample in the metering module 162is archived, the remaining fluid sample is metered and distributed tothe toxin capture and detection module 22 and the lysis and capturemodule 20. To meter and distribute a second portion of the fluid sampleto the toxin capture and detection module 22, the valves 172, 176, and177 are opened and the syringe pump 166 is turned on in a firstdirection to intake the second portion through the open valves 176 and177 into the syringe pump 166. The valves 172 and 176 are then closed,the valve 177 remains open, and the valve 178 is opened. The syringepump 166 is turned on in a second direction to force the second portionof the fluid sample from the syringe pump 166 through the open valves177 and 178 to the toxin capture and detection module 22.

To meter and distribute a third portion of the fluid sample to the lysisand capture module 20, the valves 172, 176, and 177 are opened and thesyringe pump 166 is turned on in the first direction to intake the thirdportion through the open valves 176 and 177 into the syringe pump 166.The valves 172 and 176 are then closed, the valve 177 remains open, andthe valve 180 is opened. The syringe pump 166 is turned on in the seconddirection to force the third portion of the fluid sample from thesyringe pump 166 through the open valves 177 and 180 to the lysis andcapture module 20. The syringe pump 166 is programmable to withdraw anyamount of fluid sample as is required by the application. This addsflexibility in determining how much fluid sample is provided to thetoxin capture and detection module 22 and the lysis and capture module20. In one embodiment, the second portion of fluid sample is 3 ml andthe third portion of fluid sample is 6 ml.

Although the archive module is shown in FIG. 4 as including five archivechambers, the archive module can be configured to include more or lessthan five archive modules. Further, the archiving methodology describedabove is for exemplary purposes only and any conventional methodologycan be used to purge and store subsequent fluid samples. Still further,the metering and distribution configuration and methodology describedabove in relation to FIG. 4 is but one embodiment. It is understood thatother configurations and methodologies are contemplated for metering anddistributing any number of fluid sample portions in any denomination.

FIG. 5 illustrates an exemplary schematic diagram of the toxin captureand detection module 22. The toxin capture and detection module 22includes a pump assembly including a syringe pump 222 and a distributionvalve 223, a capture module 224, and an optical detection module 234.The capture module 224 includes a capture device 228 and a reservoir226. The fluid sample provided by the distribution module 16 is receivedby the distribution valve 223 and directed to the capture module 224,where the fluid sample flows through the capture device 228. Thedistribution valve 223 is also connected to one or more reagent vesselswithin the solutions module 26.

In one embodiment, the capture device 228 is a capture chip including aplurality of pillars configured such that fluid flows around the pillarsmaking contact therewith. The pillars are prepared such that specifictoxins within the fluid sample adhere to the surface of the pillars asthe fluid flows past. The fluid sample flows through the capture chip228 and outputs the capture module 224 to waste, while any of thespecific toxins present in the fluid sample remain in the capture chip228. In one embodiment, each pillar is pre-coated with a particularantibody. Each antibody adheres to a particular type of toxin. When thefluid sample flows past the pillars, the specific toxin present withinthe fluid sample adheres to the antibody on the pillars. An example ofthe capture chip 228 is described in U.S. Pat. No. 5,707,799 and U.S.Pat. No. 5,952,173, which are both hereby incorporated by reference.

In alternative embodiments, the pillars are pre-coated with more thanone type of antibody such that each capture chip captures more than onedifferent type of toxin. More than one capture chip can be coupled inseries or in parallel to further diversify and expand the differenttypes of toxins collected. For example, a first capture chip in asequence is pre-coated with a first antibody, a second capture chip inthe sequence is pre-coated with a second antibody, and so on for as manycapture chips in the series. Additionally, one, some, or all of thecapture chips in the series can be pre-coated with more than oneantibody. For example, a capture chip can be pre-coated with multipleantibodies. Each antibody to adhere to a specific type of toxin. Thedifferent captured toxins can then be distinguished according to adistinguishing characteristic, such as different optical wavelengths. Ina series configuration, the fluid sample flows in series from the firstcapture chip to the second capture chip and so on. Although the capturedevice 228 is described above as a capture chip, the capture device 228can be any conventional device capable of capturing one or more toxins.

The toxin capture and detection module 22 includes the optical detector234 coupled to the capture device 228. The capture device 228 isconfigured such that the toxin captured within is optically accessibleto the optical detector 234. In one embodiment, the capture device 228includes an optically transparent lid. Alternatively, the captured toxinis eluted from the capture device 228 and collected in a separatecollection means, such as a vessel or reservoir. Optical detection canthen be performed on the eluted toxin in the collection means.

In this embodiment, the optical detector 234 includes a light source236, such as an LED or a laser, an optical pathway 238, such as one ormore lenses, filters and beam splitters, a fiber optics 240, and anoptical sensor 242. The optical detector 234 is configured to directlight onto the capture device 228, and to collect and measurecharacteristics of the light reflected back. The characteristics of thereflected light are used to identify the toxin(s) captured in thecapture device 228. The configuration of the optical detector 234 shownin FIG. 5 is for exemplary purposes only. In some embodiments, theoptical detector 234 is configured to include a light source, an opticalpathway to direct the light onto a specific location of the capturedevice 228 and to direct the reflected light from the capture device 228to an optical detector, and the optical detector. In other embodiments,a light source is not included. In such cases, light is emitted from thecaptured toxins, such as by chemi-luminescence. The emitted light isdetected by the optical sensor. In one embodiment, the optical detectoris any conventional optical detection device capable of measuring one ormore disparate wavelengths. The measured characteristics are providedfrom the optical detector 234 to the control module 12 for analysis.

In some embodiments, a toxin captured in the capture device 228 isidentified by forming a sandwich assay, including a flourescent marker,and then detecting the flourescent marker. The flourescent marker isoptically detectable using the optical detector 234. Each type of toxinis associated with a specific type of flourescent marker. It isunderstood that other conventional means for marking and identifying thetoxin can be used.

Once the captured toxins are interrogated by the optical detector 234,the capture device 228 is washed using washing solutions provided fromthe solutions module 26 and directed to the capture device 228. Thewashing solutions are received from the solutions module 26 by thedistribution valve 223.

Where the capture device 228 comprises multiple capture devices coupledin series, each device in series is coupled to a corresponding opticaldetector of the type described above.

FIG. 6 illustrates an exemplary schematic diagram of the lysis andcapture module 20. The lysis and capture module 20 is configured to lysecells present in the fluid sample, and to capture the nucleic acids ofthe lysed cells. The lysis and capture module 20 includes a lysischamber 260, a mixing chamber 262, a peristaltic pump 264, a pumpassembly 266 including a syringe pump 268 and a distribution valve 270,a pump assembly 272 including a syringe pump 274 and a distributionvalve 276, a purification device 278, a cooling element 280, such as athermal electric cooler, and valves 196-213. The microfluidic circuitryincluding the peristaltic pump 264, the pump assembly 266, the pumpassembly 272, and the valves 196-213 are configured to direct the fluidsample through the lysis and capture module 20, as well as to direct thevarious solutions used in processing and decontamination. The mixingchamber 262 is configured for mixing and holding solutions. For example,in some applications, one or more additional solutions are added to thefluid sample prior to lysing, and/or one or more additional solutionsare added after lysing.

The peristaltic pump 264 is configured to pressurize either the lysischamber 260, which forces fluid from the lysis chamber 260 to the mixingchamber 262, or to pressurize the mixing chamber 262, which forces fluidfrom the mixing chamber 262 to the lysis chamber 260. During eitheroperation, the appropriate valves are opened to enable such fluid flow.

The fluid sample provided by the distribution module 16 is directed tothe lysis chamber 260. In one embodiment, lysis is performed usingsonication. In some embodiments, selective lysis is performed wherespecific types of cells are lysed at different sonication energies. Inthis embodiment, the lysis and capture module 20 is configured toselectively lyse a specific type of cell at a corresponding sonicationenergy. The lysed cells are then separated from the fluid sample.Additional sonication steps can be performed on the remaining fluidsample to selectively lyse one or more additional cell types. Anexemplary apparatus and method for performing such a selective lysisprocess is described in the co-pending and co-owned U.S. patentapplication Ser. No. 10/943,601, filed on Sep. 17, 2004, and entitled“Microfluidic Differential Extraction Cartridge,” which is herebyincorporated in its entirety by reference. Alternatively, otherconventional lysis methods are utilized, such as heating and/or chemicaltreatment.

The pump assembly 266 is configured to direct the lysed fluid samplethrough the cooling element 280 and the purification device 278 to wastevia the valve 204. Nucleic acid within the lysate is purified andconcentrated as the lysate flows through the purification device 278.

In one embodiment, the purification device 278 is a purification chipincluding a plurality of pillars configured such that fluid flows aroundthe pillars making contact therewith. Nucleic acid is known to beattracted to silicon. In one embodiment, the pillars within thepurification chip are comprised of silicon such that as the fluid flowspast the pillars, nucleic acid within the fluid adheres to the pillars.Alternatively, the pillars are comprised of a material other thansilicon and are coated with silicon. Still alternatively, the pillarsare comprised of or coated with a material to which nucleic acidadheres. The fluid sample flows through the purification chip 278 andoutputs the lysis and capture module 20 to waste, while nucleic acidpresent in the fluid sample remains in the purification chip 278. Anexample of the purification chip 278 is also described in U.S. Pat. No.5,707,799 and U.S. Pat. No. 5,952,173. More than one purification chip278 can be coupled in series or in parallel. In a series configurationfor example, the fluid sample flows from a first purification chip inthe series to a second purification chip and so on. Although thepurification device 278 is described above as a purification chip, thepurification device 278 can be any conventional device capable ofcapturing nucleic acid.

The pump assembly 266 is also configured to direct a wash solutionthrough the purification device 278 to remove residual fluid samplesolution. The wash solution is provided from the solutions module 24 viathe distribution valve 270 and is directed to waste via the valve 84.Air is then blown through the purification device 228 to remove residualwash solution. The captured nucleic acids are removed from thepurification device 278 using an elution buffer. The pump assembly 272is configured to direct the elution buffer from the solutions module 24through the purification device 278 to elute the nucleic acid. Apurified and concentrated nucleic acid solution is output from thepurification device 278 and output from the lysis and capture module 20via the valve 213. In one embodiment, a heating element (not shown) iscoupled to the purification device 278. Prior to eluting the nucleicacid from the purification device 278, the heating element heats thepurification device 278, which facilitates the elution process.

The lysis and capture module 20 is also configured to back-flush thepurification device 278, either to un-block the device or as part ofwash and decontamination process. The microfluidic circuitry isconfigured to direct wash solution backwards through the purificationdevice 278 and out to waste via the valve 210.

FIG. 7 illustrates an exemplary schematic diagram of the metering andthermal cycling module 30. The metering and thermal cycling module 30 isconfigured to preamplify and amplify any nucleic acid present in thenucleic acid solution provided by the lysis and capture module 20. Themetering and thermal cycling module 30 is also configured to tag one ormore specific types of nucleic acids if present within the amplifiednucleic acid solution. The one or more specific acids are tagged using aconjugated antibody solution including a different flourescent markerfor each specific nucleic acid. The metering and thermal cycling module30 includes a plurality of solution reservoirs 321-325, a holdingreservoir 319, a metering reservoir 320, a plurality of valves 280-317,a peristaltic pump 318, a plurality of thermal cycling chambers 331-335,and a plurality of mixing reservoirs 326-330.

Each of the plurality of solution reservoirs 321-325 are coupled to thesolutions module 32 and are configured to store a specific amount ofmaster mix solution received from the solutions module 32. The holdingreservoir 319 is configured to store the nucleic acid solution outputfrom the lysis and capture module 20. The metering reservoir 320 isconfigured to meter and to store a specific amount of the nucleic acidsolution from the holding reservoir 319. In one embodiment, each of thesolution reservoirs 321-325 are configured to store 15 ul, and themetering reservoir is configured to store 10 ul. A first metered portionof the nucleic acid solution is directed from the fluid meteringreservoir 320 to the mixing reservoir 326, and the specific amount ofmixing solution from the holding reservoir 325 is directed to the mixingreservoir 326. A second portion of the nucleic acid solution is thenmetered and stored in the metering reservoir 320. The second meteredportion is directed from the metering reservoir 320 to the mixingreservoir 327, and the specific amount of mixing solution from theholding reservoir 324 is directed to the mixing reservoir 327. A meteredportion of the nucleic acid solution and a specified amount of themixing solution is provided to each of the remaining mixing reservoirs328-330 in a similar manner.

The mixed solution in the mixing reservoir 326 is directed to thethermal cycling chamber 331, the mixed solution in the mixing reservoir327 is directed to the thermal cycling chamber 332, the mixed solutionin the mixing reservoir 328 is directed to the thermal cycling chamber333, the mixed solution in the mixing reservoir 329 is directed to thethermal cycling chamber 334, and the mixed solution in the mixingreservoir 330 is directed to the thermal cycling chamber 335. A heatingelement (not shown) is coupled to each of the thermal cycling chambersto perform a thermal cycling process. In one embodiment, the thermalcycling chambers 331-335 are configured as elongated tubes capped at aeach end by a valve, and the tubes are coupled to a heating mesh to forma heating and tube assembly. An example of such a heating and tubeassembly is described in the co-owned and co-pending U.S. patentapplication Ser. No. 11/201,615, filed on Aug. 10, 2005, and entitled“Disposable Integrated Heater and Tube Assembly for Thermally-drivenChemical Reactions,” which is hereby incorporated by reference.

The microfluidic circuitry within the metering and thermal cyclingmodule 30 is configured such that multiple different thermal cyclingprocesses can be performed. After a first thermal cycling process isperformed on a first mixed solution, as described above, the resultingsolutions in the thermal cycling chambers 331-335 are back-flushed intothe corresponding mixing reservoirs 326-330. Alternatively, additionalmicrofluidic circuitry is provided which directs solutions from thethermal cycling chambers 331-335 to their respective mixing reservoirs326-330. Additional mixing solutions can be provided to the mixingreservoirs 326-330 from the solution reservoirs 321-325. The mixingsolutions provided during this step can be the same or different thanthe mixing solutions provided during the first thermal cycling process.The mixed solutions are then directed back to the thermal cyclingchambers 331-335 for a second thermal cycling process. Additionalthermal cycling processes can be performed in this manner. In oneapplication, a pre-amplification process is performed during the firstthermal cycling process and an amplification process is performed duringthe second thermal cycling process. An example of one suchpre-amplification and amplification process is described in theco-pending, co-owned U.S. Patent Application, Serial No. (MFSI 00600),which is hereby incorporated by reference. The amplification processresults in an amplified nucleic acid solution. The amplified nucleicacid solution is output from the metering and thermal cycling module 30.

One or more additional processing steps can be performed on theamplified nucleic acid solution prior to being output from the meteringand thermal cycling module 30. Such additional processing steps preparethe amplified nucleic acid solution for interrogation by the opticaldetection module 34. The amplified nucleic acid solution is back-flushedfrom the thermal cycling chambers 331-335 to the corresponding mixingreservoirs 326-330. An additional solution is added to each of themixing reservoirs. The additional solution is configured to adhere toone or more specific types of nucleic acids if present within theamplified nucleic acid solution. The resulting product includes adifferent flourescent marker for each specific nucleic acid. Thisproduct is then output from the metering and thermal cycling module 30.It is understood that alternative chemistries can be used to detect thepresence of the specific types of nucleic acids.

Although the metering and thermal cycling module 30 shown in FIG. 7 isconfigured with five thermal cycling chambers, five mixing reservoirs,and five solution reservoirs, the metering and thermal cycling module 30can be configured with more or less than five thermal cycling chambers,five mixing reservoirs, and five solution reservoirs. Stillalternatively, an alternative mixing method eliminates the mixingreservoirs and relies on mixing within the fluid lines themselves duringtransport of the fluids from the solutions reservoirs to the thermalcycling chambers.

FIG. 8 illustrates an exemplary schematic diagram of the opticaldetection module 34. The optical detection module 34 includes a pumpassembly 340 including a syringe pump 341 and a distribution valve 342,a fluid line 344 including an interrogation channel 343, and an opticaldetector 346. The fluid line 344 receives the amplified nucleic acidsolution output from the metering and thermal cycling module 30. Theinterrogation channel 343 is an optically transparent portion of thefluid line 344 that enables optical analysis to be performed by theoptical detector 346 as the amplified nucleic acid solution passesthrough the optically transparent portion. In one embodiment, theinterrogation channel 343 is integrated within the microfluidiccircuitry connecting the metering and thermal cycling module 30 to thewaste module 28 (FIG. 2). In this configuration, optical measurementsare taken of the amplified nucleic acid solution as the solution isdirected to waste. Alternatively, a collection vessel is coupled to thefluid line 344, and the amplified nucleic acid solution is collected inthe collection vessel, where optical measurements are taken.

The optical detector 346 includes a light source 348, such as awhite-light LED or a laser, an optical pathway 350, such as one or morelenses, filters and beam splitters, a fiber optics 352, and an opticalsensor 354. The optical detector 346 is functionally equivalent to theoptical detector 234 (FIG. 5) in the toxin capture and detect module 22.The optical detector 346 is configured to direct light into theinterrogation channel 343, and to collect and measure characteristics ofthe light reflected back. The characteristics of the reflected light areused to determine if specific types of nucleic acids are present in theamplified nucleic acid solution. The configuration of the opticaldetector 346 shown in FIG. 8 is for exemplary purposes only. In someembodiments, the optical detector 346 is configured to include a lightsource, an optical pathway to direct the light onto the interrogationchannel 343 and to direct the reflected light from the interrogationchannel 343 to an optical detector, and the optical detector. In otherembodiments, a light source is not included. In such cases, light isemitted from the captured toxins, such as by chemi-luminescence. Theemitted light is detected by the optical sensor. In one embodiment, theoptical detector is any conventional optical detection device capable ofmeasuring one or more disparate wavelengths. The measuredcharacteristics are provided from the optical detector 346 to thecontrol module 12 for analysis.

The particle collection and detection system 10 is a fully integratedand automated system configured to detect the presence of specificairborne particles. FIG. 9 illustrates an exemplary automated processperformed by the particle collection and detection system 10. At thestep 400, intake ambient air into the air collection module 14. Air iscontinuously taking in by the air collection module 14 throughout theentire process. At the step 405, periodically output a fluid sample fromthe air collection module 14 according to a defined schedule. The outputfluid sample includes airborne particles collected from the ambient air.At the step 410, meter and distribute the fluid sample. At the step 415,archive a first portion of the fluid sample. At the step 420, capture,purify and concentrate toxins from within a second portion of the fluidsample. At the step 425, determine the presence of toxins captured inthe step 420. In one embodiment, optical detection is used to detect thepresence of toxins.

At the step 430, lyse cells in a third portion of the fluid sample. Thisgenerates a lysate solution. At the step 435, meter and distribute thelysate solution. At the step 440, perform a pre-amplification process oneach metered portion of the first lysate. At the step 445, perform anamplification process on each metered portion of the first lysate togenerate an amplified nucleic acid solution. The pre-amplificationprocess and the amplification process include thermal cycling. At thestep 450, determine the presence of one or more specific types ofnucleic acids in the amplified nucleic acid solution and identifying theone or more specific types of nucleic acids. The steps 430 through 450are performed in parallel with the steps 420 through 425, therebysimultaneously processing the fluid sample.

At the step 455, generate an alarm signal if one or more toxins aredetermined at the step 425 or one or more specific nucleic acids aredetermined at the step 450. At the step 460, reset the system to processthe next fluid sample to be output by the air collection module 14. Thesystem is reset by decontaminating the microfluidic circuitry throughwhich the fluid sample passed, any fluid sample collection vessels, thecapture devices used to capture the toxins, the purification devicesused to purify the nucleic acids, any purged archive chambers, and thethermal cycling chambers. Decontamination is performed using anyconventional rinsing and washing steps. After the system is reset, andat the next scheduled interval, the next fluid sample is output from theair collection module 14 and processed as described above. This processis continuously repeated for successive fluid samples. The particlecollection and detection system functions independently, or is networkedto a remote monitoring and/or control location to which measuredcharacteristics and/or post-analysis results are transmitted and/or fromwhich control signals are received.

In an exemplary application, the collection and detection system 10operates continuously 24 hours a day, 7 days a week. Every three hoursthe air collection module outputs a 10 ml fluid sample to thedistribution module 16. 1 ml of the 10 ml fluid sample is metered anddistributed to the archive module 18, 3 ml to the toxin capture anddetection module 22, and 6 ml to the lysis and capture module 20. Thelysis and capture module 20 outputs a 50 ul sample for each 6 ml inputsample. The metering and thermal cycling module 30 receives as input the50 ul sample from the lysis and capture module 20 and 15 ul aliquotsfrom the solutions module 32. The metering and thermal cycling module 30outputs five, 25 ul samples for each 50 ul input sample received fromthe lysis and capture module 20. Each of the five, 25 ul samples areanalyzed by the optical detection module 34. The above timing, samplesizes, and distribution ratios are for exemplary purposes only. Thespecific timing, sample sizes, and distribution ratios are applicationspecific and the collection and detection system 10 is configuredaccordingly. Positive and negative control samples can be substitutedfor one or more of the 25 ul fluid samples processed by the metering andthermal cycling module 30, thereby verifying the accuracy of theanalysis performed on any given input fluid sample.

A second embodiment of a collection and detection system is directed toa detect to warn system in which the presence of specific types ofparticles are detected, and may or may not be identified. FIG. 10illustrates an exemplary functional block diagram of the secondembodiment of the integrated collection and detection system. Theintegrated collection and detection system 500 includes an aircollection module 510, a confirmation device 520, and a control module530. Fluid is directed between the fluid interface 514 and theconfirmation device 520, and within the confirmation device 520, usingmicrofluidic circuitry.

The air collection module 510 is configured to intake ambient air,detect the presence of one or more different types of airborne particleswithin the ambient air, and collect the airborne particles, such aswithin a fluid. The air collection module 510 includes a triggeringmechanism 512 and a fluid interface 514. The fluid interface 514 isconfigured to receive ambient air, including airborne particles presenttherein, that is drawn into the collection and detection system 500 andto collect the airborne particles into a fluid solution, also referredto as a fluid sample. The fluid interface 500 includes a fan to generateairflow into the collection and detection system 500. In someembodiments, the airborne particles are collected by eluting particlescollected on the fan and then collecting the resulting fluid solutionincluding the eluted particles. One such method of collecting theairborne particles into a fluid solution is described in the co-owned,co-pending U.S. Patent Application Serial No. (MFSI-00500), entitled“Automated Particle Collection Off of Fan Blades into a Liquid Buffer,”which is hereby incorporated by reference. The fluid solution can bestored in a collection vessel within the fluid interface 514, or in acollection vessel external to the fluid interface 514 and/or the aircollection module 510.

The triggering mechanism 514 is positioned to continuously monitor theairflow, and the airborne particles within the airflow, directed to thefluid interface 514. The triggering mechanism 512 includes a lightsource, such as a laser or a white-light LED, to generate a light beamthat is directed at the airflow. The light beam impinges the airborneparticles within the airflow. The triggering mechanism 512 also includesa light collector, such as an optical sensor, to measure one or moreoptical characteristics associated with the light after impinging theairborne particles. In some embodiments, the wavelength of the lightreflected off the airborne particles is measured. The triggeringmechanism 512 is non-destructive in relation to the airborne particles.

The optical characteristics measured by the triggering mechanism 512 areprovided to the control module 530. The optical characteristics arecompared to known optical characteristics by the control module 530 todetermine if one or more different types of specific biologicalparticles are present in the airflow. If it is determined that one ormore different types of specific biological particles are present, thana trigger signal is generated by the control module 530. Alternatively,the triggering mechanism 512 includes logic circuitry to determine ifone or more different types of specific biological particles are presentand to generate the trigger signal, if necessary. Still alternatively,the triggering mechanism 512 includes logic circuitry to determine ifone or more different types of specific biological particles arepresent, and the control module 530 generates the trigger signal, ifnecessary.

In response to the trigger signal, the fluid sample, or a portionthereof, is directed to the confirmation device 520 to confirm thepresence of the one or more different types of specific biologicalparticles. The confirmation device 520 includes a solutions module 522and a toxin capture and detection module 524.

The toxin capture and detection module 524 of the second embodiment isphysically and operationally equivalent to the toxin capture anddetection module 22 of the first embodiment with the exception that theone or more capture devices and the optical detection module within thetoxin capture and detection module 524 are configured to capture anddetect specific pathogens in addition to specific toxins. Some pathogensare detectable using immuno assay. In some embodiments, the one or morecapture devices within the toxin capture and detection module 524 arepre-coated with one or more specific antibodies known to adhere tospecific pathogens, in addition to the one or more specific antibodiesknown to adhere to specific toxins as described in relation to the toxincapture and detection module 22. In these embodiments, the opticaldetection module within the toxin capture and detection module 524 isconfigured to measure one or more optical characteristics of anycaptured toxin or pathogen, which are used to determine the presence ofeach of the specific antibodies.

The raw data obtained by the toxin capture and detection module 524,such as the measured optical characteristics, is provided to the controlmodule 530, where it is used to determine the presence and identity ofone or more specific types of toxins and/or pathogens. If a specifictoxin or pathogen is detected, the control module 530 generates an alarmsignal. Alternatively, the raw data collected by the toxin capture anddetection module 524 is sent to a remote location, such as the centralmonitoring point 50 (FIG. 1) for analysis.

The solutions module 522 is similar to the solutions module 26 (FIG. 2)in that it provides solutions used during the capture steps performed inthe toxin capture and detection module 524. For example, the solutionsmodule 522 includes wash solutions and antibody solutions.

The collection and detection system 500 is configured to be re-used suchthat ambient air is continuously interrogated and successive fluidsamples output by the air collection module 510 are processed. As such,the toxins capture and detection module 524 and all interconnectingmicrofluidic circuitry are decontaminated between cycles. Varioussolutions are used to perform the rinse and wash steps duringdecontamination, these solutions are included in the solutions module522.

The control module 530 is coupled to each module to control operation ofthe collection and detection system 500. Such control enables completeautomation of the collection and detection process, without need ofhuman intervention. The control module 530 is also configured to analyzethe raw data provided by the toxin capture and detection module 524 andto generate any appropriate alarm or trigger signals. In response to analarm signal, the control module 530 initiates a localized audio and/orvisual alarm and/or transmits a notification signal to a networked localmonitoring location or a centralized monitoring location.

The analyzed fluid samples, elution buffers, mixing solutions, rinses,washes, purged archive samples, and other solutions related to theprocessing of fluid samples and subsequent decontamination of thecollection and detection system 500 are directed to a waste module (notshown). Alternatively, fluid samples analyzed and subsequently output bythe toxin capture and detection module 524 can be archived, either in alocal or a remote storage vessel.

FIG. 11 illustrates an exemplary automated process performed by theparticle collection and detection system 500. At the step 540, intakeambient air into the air collection module 510. Air is continuouslytaken in by the air collection module 510 throughout the entire process.At the step 545, airborne particles within the ambient air areinterrogated to measure one or more optical characteristics associatedwith the airborne particles. In some embodiments, a laser beam is usedto interrogate the airborne particles such that the wavelengths of lightreflected from the laser beam impinging the airborne particles ismeasured. At the step 550, the measured optical characteristics arecompared to known optical characteristics associated with one or moredifferent types of biological particles. If it is determined that thereis not a match at the step 550, then the method repeats the step 540 and545. If however it is determined that there is a match at the step 550,then at the step 555 a trigger signal is generated. Generation of thetrigger signal indicates that at least one type of biological particleis present within the ambient air.

At the step 560, a fluid sample is generated that includes the particlesfrom the ambient air. In response to the trigger signal, the fluidsample, or a portion thereof, is directed to the confirmation device520. The step 560 can be performed after the step 545 such that thefluid sample is always generated, regardless of a match made between themeasured optical characteristics and known optical characteristics. Thestep 560 can also be performed concurrently with the step 550, and ifnecessary the step 555. At the step 565, the confirmation device 520confirms that one or more specific types of biological particles arepresent. The biological particles are either specific types of toxins orspecific types of pathogens. In some embodiments, the confirmationdevice 520 confirms the presence of one or more different types oftoxins and/or pathogens using immuno assays. In some embodiments, theconfirmation device 520 identifies one or more of the different types oftoxins and/or pathogens. In some embodiments, the confirmation device520 generates an alarm signal if the presence of one or more differenttypes of toxins and/or pathogens is confirmed.

A third embodiment of a collection and detection system combines thefunctionality of the collection and detection system 10 of FIG. 1 andthe collection and detection system 500 of FIG. 10. In this thirdembodiment, the collection and detection system 500 is adapted toperform a first level of detection in which the presence of one or moretoxins and/or pathogens are detected, and upon such detection, thecollection and detection system 10 is adapted to perform a second levelof detection in which the one or more toxins and/or pathogens areidentified.

FIG. 12 illustrates an exemplary functional block diagram of the thirdembodiment of the integrated collection and detection system. Theintegrated collection and detection system 600 includes the aircollection module 510 and the confirmation device 520 of the collectionand detection system 500, and the distribution module 16, the archivemodule 18, the lysis and capture module 20, the toxin capture anddetection module 22, the solutions module 24, the solutions module 26,the waste module 28, the metering and thermal cycling module 30, thesolutions module 32, and the optical detection module 34 of thecollection and detection system 10. The collection and detection system600 also includes a distribution module 610 and a control module 620.Each of the modules are fluidically coupled as appropriate to directfluid sample and solutions within the collection and detection system600.

Control of the collection and detection system 600 is maintained by thecontrol module 620, which includes the functionality of the controlmodule 12 of the collection and detection system 10 and the controlmodule 530 of the collection and detection system 500. Alternatively,control is distributed locally, such as by adding the control module 530to control the first level of detection and by adding the control module12 to control the second level of detection. Such local control modulescommunicate with each other to coordinate their respective functions.Still alternatively, control is distributed locally, such as by addingthe control module 530 and the control module 12, and maintaininghigh-level control over the collection and detection system 600 by aglobal control module coupled to the local control modules. The controlmodule 620 is coupled to each of the modules in the collection anddetection system 600.

The distribution module 610 is configured to receive the fluid sampleoutput from the fluid interface 514. The distribution module 610includes microfluidic circuitry and storage vessels. The fluid samplereceived from the fluid interface 514 is metered and distributedaccording to predetermined ratios. A first portion of the fluid sampleis metered and distributed to the confirmation device 520 in response tothe trigger signal. The remaining portion of the fluid sample remainsstored in the distribution module 610. If the confirmation device 520confirms the presence of one or more specific types of biologicalparticles, then the alarm signal is generated. In response to the alarmsignal, the remaining portion of the fluid sample is distributed fromthe distribution module 610 to the distribution module 16. The fluidsample is then processed by the toxin capture and detection module 22,the lysis and capture module 20, the metering and thermal cycling module20, and the optical detection module 34 to identify particles within thefluid sample. In some embodiments, a single distribution module can beconfigured to combine the functionality of the distribution module 610and the distribution module 16.

If the triggering mechanism 512 does not generate a trigger signal, thefluid sample is stored in the distribution module 610 until the nextscheduled interval for providing the fluid sample to the distributionmodule 16 to process. If the triggering mechanism 512 does generate atrigger signal but the confirmation device 520 does not generate analarm signal, the remaining fluid sample is stored in the distributionmodule 610 until the next scheduled interval. Alternatively, if thetriggering mechanism 512 does generate a trigger signal, the remainingfluid sample is distributed to the distribution module 16 to processwhether or not the confirmation device 520 generates an alarm signal.The fluid interface 514 continues to output fluid sample to be stored inthe distribution module 610 regardless of whether or not the triggersignal or alarm signal are generated.

In operation of the collection and detection system 600, the triggeringmechanism 512 and the confirmation device 520 perform a first level ofdetection that determines if specific types of biological particles arepresent in the ambient air. If the first level of detection confirms thepresence of one or more specific types of biological particles, a secondlevel of detection is performed by the toxin capture and detectionmodule 22, the lysis and capture module 20, the metering and thermalcycling module 20, and the optical detection module 34. The second levelof detection identifies one or more specific toxins and/or one or morespecific types pathogens.

FIG. 13 illustrates an exemplary automated process performed by thethird embodiment of the particle collection and detection system. At thestep 625, intake ambient air into the air collection module 510. Air iscontinuously taken in by the air collection module 510 throughout theentire process. At the step 630, airborne particles within the ambientair are interrogated to measure one or more optical characteristicsassociated with the airborne particles. In some embodiments, a laserbeam is used to interrogate the airborne particles such that thewavelengths of light reflected from the laser beam impinging theairborne particles is measured. At the step 635, a fluid sample isgenerated that includes the particles from the ambient air. At the step640, the measured optical characteristics are compared to known opticalcharacteristics associated with one or more different types ofbiological particles. If it is determined that there is not a match atthe step 640, then the method returns to the step 625. If however it isdetermined that there is a match at the step 640, then at the step 645 atrigger signal is generated. Generation of the trigger signal indicatesthat at least one type of biological particle is detected within theambient air.

In response to the trigger signal, at the step 650 a first portion ofthe fluid sample is metered and distributed to the confirmation device520. At the step 655, the confirmation device 520 confirms that one ormore specific types of biological particles are present in the firstportion of the fluid sample. The biological particles are eitherspecific types of toxins or specific types of pathogens. In someembodiments, the confirmation device 520 confirms the presence of one ormore different types of toxins and/or pathogens using immuno assays. Insome embodiments, the confirmation device 520 identifies one or more ofthe different types of toxins and/or pathogens. If it is determined atthe step 655 that the one or more specific types of biological particlesare not present in the first portion of the fluid sample, then themethod returns to the step 625. If however it is determined at the step655 that the one or more specific types of biological particles arepresent in the first portion of the fluid sample, then at the step 660 afirst alarm signal is generated. Generation of the first alarm signalindicates that at least one type of biological particle is detectedwithin the first portion of the fluid sample.

At the step 665, a remaining portion of the fluid sample is metered anddistributed to the archive module 18, the toxin capture and detectionmodule 22, and the lysis and capture module 20. At the step 670, secondportion of the fluid sample is archived. At the step 675, toxins fromwithin a third portion of the fluid sample are captured, purified andconcentrated. At the step 680, the presence of toxins captured in thestep 675 is determined and the toxins are identified. In one embodiment,optical detection is used to detect and identify the toxins.

At the step 685, cells in a fourth portion of the fluid sample arelysed. This generates a lysate solution. At the step 690, the lysatesolution is metered and distributed. At the step 695, apre-amplification process is performed on each metered portion of thefirst lysate. At the step 700, an amplification process is performed oneach metered portion of the first lysate to generate an amplifiednucleic acid solution. The pre-amplification process and theamplification process include thermal cycling. At the step 705, thepresence of one or more specific types of nucleic acids in the amplifiednucleic acid solution is determined and the one or more specific typesof nucleic acids are identified. The steps 685 through 705 are performedin parallel with the steps 675 through 680, thereby simultaneouslyprocessing the fluid sample.

At the step 710, a second alarm signal is generated if one or moretoxins are determined at the step 680 or one or more specific nucleicacids are determined at the step 705. At the step 715, the system isreset in order to process the next fluid sample to be output by the aircollection module 14. The system is reset by decontaminating themicrofluidic circuitry through which the fluid sample passed, any fluidsample collection vessels, the capture devices used to capture thetoxins, the purification devices used to purify the nucleic acids, anypurged archive chambers, and the thermal cycling chambers.Decontamination is performed using any conventional rinsing and washingsteps.

Embodiments of the integrated particle collection and detection systemare described above in relation to a bio-threat application. It isunderstood that the integrated particle collection and detection systemcan also be used to collect non-harmful air particles and in general theintegrated particle collection and detection system can be used tocollect and analyze any airborne particles.

The network configuration described in relation to FIG. 1 includes thefirst embodiment of the collection and detection system, the collectionand detection system 10. It is understood that one, some, or all of theembodiments of the collection and detection system, for example thecollection and detection system 10, the detection and collection system500, and the collection and detection system 600, can be networked in asimilar manner and in any combination.

The embodiments of the collection and detection system described aboveare for exemplary purposes. The microfluidic circuitry and module natureof the integrated collection and detection system provides flexibilityand extensibility to interconnect and configure the modules, andassociated sub-modular components, into any desired combination.Additionally, the specific configurations described for each of themodules is for exemplary purposes. The microfluidic circuitry andconstituent components of each module can be adapted into any number ofconfigurations to perform the described functionality.

The present invention has been described in terms of specificembodiments incorporating details to facilitate the understanding of theprinciples of construction and operation of the invention. The specificconfigurations shown and the methodologies described in relation to theair collection device, the confirmation device, the distribution module,the archive module, the lysis and capture module, the toxin capture anddetection module, the metering and thermal cycling module, the opticaldetection module, the control module, and the interconnectionstherebetween are for exemplary purposes only. Such reference herein tospecific embodiments and details thereof is not intended to limit thescope of the claims appended hereto. It will be apparent to thoseskilled in the art that modifications may be made in the embodimentchosen for illustration without departing from the spirit and scope ofthe invention.

1. A detection apparatus to detect the presence of one or more differenttypes of particles, the detection apparatus comprises: a. a first leveldetection device configured to interrogate ambient air for the presenceof one or more different types of airborne biological particles, whereinthe first level detection device is further configured to generate atrigger signal in response to detecting one or more of the differenttypes of airborne biological particles; b. a fluid interface configuredto receive airborne particles within the ambient air and to output afluid sample including the particles; and c. a second level detectiondevice configured to receive the fluid sample in response to the triggersignal and to confirm the presence of the one or more different types ofbiological particles using an immuno assay.
 2. The detection apparatusof claim 1 further comprising microfluidic circuitry to couple the fluidinterface to the second level detection device.
 3. The detectionapparatus of claim 2 wherein the microfluidic circuitry includes acollection vessel to store the fluid sample.
 4. The detection apparatusof claim 3 wherein the microfluidic circuitry is configured to meter thefluid sample into one or more portions and to distribute a first portionof the fluid sample to the second level detection device.
 5. Thedetection apparatus of claim 1 wherein the second level detection deviceis configured to identify the one or more different types of biologicparticles using the immuno assay.
 6. The detection apparatus of claim 1wherein the biologic particle comprises a pathogen.
 7. The detectionapparatus of claim 1 wherein the biological particle comprises a toxin.8. The detection apparatus of claim 1 wherein the second level detectiondevice comprises one or more capture devices each configured to captureone or more different types of biological particles from the fluidsample.
 9. The detection apparatus of claim 8 wherein the second leveldetection device further comprises an optical detection moduleconfigured to optically detect the presence of the captured one or moredifferent types of biological particles within the one or more capturedevices.
 10. The detection apparatus of claim 1 further comprising acontrol module configured to control the operation of the first leveldetection device, the fluid interface, and the second level detectiondevice to enable the detection apparatus to function automatically. 11.The detection apparatus of claim 1 wherein the detection apparatus isconfigured to confirm the presence of the one or more different types ofbiological particles within about 5 minutes from generating the triggersignal.
 12. The detection apparatus of claim 1 wherein the first leveldetection device comprises an air collection device configured to intakethe ambient air to be interrogated.
 13. The detection apparatus of claim12 wherein the first level detection device further comprises atriggering mechanism configured to monitor the ambient air taken in bythe air collection device and to generate the trigger signal.
 14. Thedetection apparatus of claim 13 wherein the triggering mechanismcomprises a light source configured to direct light onto the airborneparticles and a light collector configured to measure one or moreoptical characteristics of light impinging the airborne particles. 15.An autonomously functioning detection apparatus to detect the presenceof one or more different types of particles, the autonomouslyfunctioning detection apparatus comprises: a. a first level detectiondevice configured to automatically interrogate ambient air for thepresence of one or more different types of airborne biologicalparticles, wherein the first level detection device is furtherconfigured to automatically generate a trigger signal in response todetecting one or more of the different types of airborne biologicalparticles; b. a fluid interface configured to automatically receiveairborne particles within the ambient air and to automatically output afluid sample including the particles; c. a second level detection deviceconfigured to automatically receive the fluid sample in response to thetrigger signal and to automatically confirm the presence of the one ormore different types of biological particles; and d. a control moduleconfigured to provide control signals to the first level detectiondevice, the fluid interface, and the second level detection device toenable the detection apparatus to function autonomously.
 16. Theautonomously functioning detection apparatus of claim 15 wherein thesecond level detection device is configured to automatically confirm thepresence of the one or more different types of biological particlesusing an immuno assay.
 17. The autonomously functioning detectionapparatus of claim 16 wherein the second level detection device isconfigured to automatically identify the one or more different types ofbiologic particles using the immuno assay.
 18. The autonomouslyfunctioning detection apparatus of claim 15 further comprisingmicrofluidic circuitry to couple the fluid interface to the second leveldetection device.
 19. The autonomously functioning detection apparatusof claim 18 wherein the microfluidic circuitry includes a collectionvessel to automatically store the fluid sample.
 20. The autonomouslyfunctioning detection apparatus of claim 19 wherein the microfluidiccircuitry is configured to automatically meter the fluid sample into oneor more portions and to automatically distribute a first portion of thefluid sample to the second level detection device.
 21. The autonomouslyfunctioning detection apparatus of claim 15 wherein the biologicparticle comprises a pathogen.
 22. The autonomously functioningdetection apparatus of claim 15 wherein the biological particlecomprises a toxin.
 23. The autonomously functioning detection apparatusof claim 15 wherein the second level detection device comprises one ormore purification devices each configured to automatically capture oneor more different types of biological particles from the fluid sample.24. The autonomously functioning detection apparatus of claim 23 whereinthe second level detection device further comprises an optical detectionmodule configured to optically detect the presence of the captured oneor more different types of biological particles within the one or morepurification devices.
 25. The autonomously functioning detectionapparatus of claim 15 wherein the autonomously functioning detectionapparatus is configured to automatically confirm the presence of the oneor more different types of biological particles within about 5 minutesfrom generating the trigger signal.
 26. The autonomously functioningdetection apparatus of claim 15 wherein the first level detection devicecomprises an air collection device configured to automatically intakethe ambient air to be interrogated.
 27. The autonomously functioningdetection apparatus of claim 26 wherein the first level detection devicefurther comprises a triggering mechanism configured to automaticallymonitor the ambient air taken in by the air collection device and toautomatically generate the trigger signal.
 28. The autonomouslyfunctioning detection apparatus of claim 27 wherein the triggeringmechanism comprises a light source configured to automatically directlight onto the airborne particles and a light collector configured toautomatically measure one or more optical characteristics of lightimpinging the airborne particles.
 29. A detection apparatus to detectthe presence of one or more different types of particles, the detectionapparatus comprises: a. an air collection device configured to intakeambient air including airborne particles; b. a triggering mechanismconfigured to interrogate the ambient air for the presence of one ormore different types of airborne biological particles and to generate atrigger signal in response to detecting one or more of the differenttypes of airborne biological particles; c. a fluid interface configuredto receive airborne particles within the ambient air and to output afluid sample including the particles; d. a confirmation deviceconfigured to receive the fluid sample in response to the trigger signaland to confirm the presence of the one or more different types ofbiological particles using an immuno assay; and e. a control moduleconfigured to provide control signals to the air collection device, thetriggering mechanism, the fluid interface, and the confirmation deviceto enable the detection apparatus to function autonomously.
 30. Thedetection apparatus of claim 29 further comprising microfluidiccircuitry to couple the fluid interface to the confirmation device. 31.The detection apparatus of claim 30 wherein the microfluidic circuitryincludes a collection vessel to store the fluid sample.
 32. Thedetection apparatus of claim 31 wherein the microfluidic circuitry isconfigured to meter the fluid sample into one or more portions and todistribute a first portion of the fluid sample to the confirmationdevice.
 33. The detection apparatus of claim 29 wherein the confirmationdevice is configured to identify the one or more different types ofbiologic particles using the immuno assay.
 34. The detection apparatusof claim 29 wherein the biologic particle comprises a pathogen.
 35. Thedetection apparatus of claim 29 wherein the biological particlecomprises a toxin.
 36. The detection apparatus of claim 29 wherein theconfirmation device comprises one or more purification devices eachconfigured to capture one or more different types of biologicalparticles from the fluid sample.
 37. The detection apparatus of claim 36wherein the confirmation device further comprises an optical detectionmodule configured to optically detect the presence of the captured oneor more different types of biological particles within the one or morepurification devices.
 38. The detection apparatus of claim 29 whereinthe detection apparatus is configured to confirm the presence of the oneor more different types of biological particles within about 5 minutesfrom generating the trigger signal.
 39. The detection apparatus of claim29 wherein the triggering mechanism comprises a light source configuredto direct light onto the airborne particles and a light collectorconfigured to measure one or more optical characteristics of lightimpinging the airborne particles.
 40. A method of detecting the presenceof one or more different types of biological particles, the methodcomprising: a. collecting ambient air including airborne particles; b.measuring one or more optical characteristics of the airborne particles;c. comparing the measured one or more optical characteristics to knownoptical characteristics associated with one or more types of biologicalparticles; d. generating a trigger signal if the one or more of themeasured one or more optical characteristics match one or more of theknown optical characteristics; e. converting the airborne particles to afluid sample; and f. analyzing the fluid sample according to an immunoassay to confirm the presence of the one or more specific types ofbiological particles.